OBJECTIVE:To establish a method for the content determination of loxoprofen silver in Loxoprofen silver cream, and provide reference for the quality control of the preparation. METHODS:HPLC was performed on the column of Diamonsil C18 with mobile phase of methanol-water-glacial acetic acid-triethylamine(55:45:0.1:0.1,V/V/V/V)at a flow rate of 1.0 mL/min,de-tection wavelength was 223 nm,column temperature was 30 ℃,and the injection volume was 20 μL. RESULTS:The linear range of loxoprofen silver was 6.53-130.7μg/mL(r=0.9999);the limit of quantitation was 0.253μg/mL,limit of detection was 0.076μg/mL;RSDs of precision,stability and reproducibility tests were lower than 2%;recovery was 96.79%-103.68%(RSD=2.23%, n=9). CONCLUSIONS:The method is simple,accurate and reproducible,and can be used for the content determination of Loxo-profen silver cream.

Aim To explore the effects and mechanism of ginsenoside-Rg1 on SK-N-SH cells treated with chronic morphine and naloxone-precipitated withdrawal. Methods Cells were pretreated with ginsenoside-Rg1 1,2,4,8,16,32 ?mol?L-1 for 24 h,then incubated for 24 h with morphine ( 100 ?mol?L-1 ) . MTT colorimetr was used to study the effects of ginsenoside-Rg1 on the multiplication of the cells treated with chro-nic morphine. After stimulated by the same concentra-tion of morphine,cells were added with different concentrations of Rg1 1,2,4 ?mol?L -1 for 24 h before stimulated with 10 ?mol?L -1 NAL. Fuorospectrophotometry RT-PCR and Western blot techniques were used to detect the effects of ginsenoside-Rg1 on the [Ca2+ ]i,CaMKⅡ ? mRNA and protein expression of the SK-N-SH cells treated with chronic morphine and naloxone-precipitated withdrawal. Results ① Compared with control group,morphine significantly inhibited cell multiplication and resulted in calcium overload,and the expression of CaMKⅡ-? mRNA and protein noticeably increased ( P

A capillary electrophoretic method was developed for the separation of enantiomers of naphthylgly-cidic ether(NGE). Several cyclodextrins(CDs) were applied as the chiral selectors and it was found that the ionic modified highly sulfated cyclodextrin(HS-β-CD) could give satisfactory enantioselectivity. In addition,the effects of the pH value of the buffer system,the concentration of the HS-β-CD,capillary temperature and the running voltage on the chiral separation were investigated systematically. The result showed that under the optimized conditions of pH 2. 5,20 mmol/L H_3PO_4-triethanolamine buffer containing 2% highly sulfated cyclodextrin ( HS-β-CD) at capillary temperature 20℃ in the reversed polarity voltage of - 18 kV,the enantiomers of naphthylglycidic ether were baseline separated. This method is simple,precise and can be applied to the chiral separation of naphthylglycidic ether enantiomers and determination of enantiomers excess(ee,% ).

Objective: To intensively investigate sporadic CMT patients, we have analyzed the LMNA gene in this study in a series of 32 unrelated CMT patients. Methods: Twelve exons of the LMNA gene were amplified from genetomic DNA. PCR products of each exon were analyzed by single strand conformational polymorphism (SSCP). Results: No abnormal SSCP pattern, suggesting no mutation in our CMT patients, was detected. Conclusion: The CMT diseases resulted from the mutations of LMNA gene were rare.

Objective To investigate the expressions of ezrin,E-cadherin,paxillin and integrin β1 in skin squamous cell carcinoma (SCC) and their significance.Methods The expressions of ezrin,E-cadherin,paxillin and.integrin β1 were immunohistochemically detected in tissue specimens from 30 patients with SCC and 10 normal human controls.Results The expression rates of ezrin and E-cadherin in normal human keratinocytes were significantly higher than those in SCC cells [ 100.00％ (10/10)vs.56.67％(17/30),x2 =6.42,P ＜ 0.05; 100.00％ (10/10) vs 13.33％ (4/30),x2=24.76,P ＜ 0.05],while the expression rates of paxillin and integrin β1 in normal human keratinocytes were significantly lower than those in SCC cells [0.00vs.70.00％ (21/30),x2 =14.74,P ＜ 0.01; 10.00％ (1/10) vs.66.67％ (20/30),x2 =9.66,P ＜ 0.05].In SCC cells,there was a significant positive correlation between the expression of ezrin and E-cadherin (r =0.52,P ＜0.01 ),but a negative correlation between the expression of paxillin and E-cadherin and between that of integrin β1 and E-cadherin (r =- 0.56,- 0.52 respectively,both P ＜ 0.01); no significant correlation was observed between the expression of ezrin and paxillin or integrin β1,or between the that of paxillin and integrin β1 (r =0.5,0.24 and 0 respectively,all P ＞ 0.05).Conclusions The low expression of E-cadherin and high expression of paxillin and integrin β1 may synergistically promote the invasion and metastasis of tumor cells,while integrin β1 and E-cadherin may play an antagonistic role in the metastasis of SCC.

Objective To observe the biological characteristics of db/db mice, and to provide the basis for application of db/db mice in experimental research.Methods Spontaneous type 2 diabetes BKS.Cg-Dock7m +/+ Leprdb/JNju mice and wild type mice of the same age were used in this study.Their fasting blood glucose was determined at 8,12,16, 20 and 24 weeks of age, the body weight was recorded at 10, 12,16, 20 and 24 weeks of age, the levels of serum insulin, total cholesterol and triglyceride were measured, the organ weight and liver coefficient were determined, and the liver and pancreas were taken for pathological examination at 24 weeks of age.Results The db/db mice maintained a high level of fasting blood glucose and body weight.Levels of serum insulin, total cholesterol, and triglyceride were significantly higher than the wild type mice.Compared with wild type mice, liver weight and kidney weight were also significantly increased.Obvious pathological changes of liver and pancreas were observed in 24-week old db/db mice.Conclusions db/db mouse has obvious characteristics of type 2 diabetes, such as hyperglycemia, hyperlipidemia, and hyperinsulinemia, can maintain stable levels of high blood glucose,and is an ideal animal model for experimental study of type 2 diabetes mellitus.

Aim To investigate the proliferative effect and the apoptosis of rat epididymal epithelial cells induced by podophyllotoxin and its underlying mechanisms.Methods Primary epididymal epithelial cells were cultured in vitro.CCK-8 assay was used to analyze proliferation of epididymal epithelial cells induced by podophyllotoxin on 24, 48 and 72 h.The ultra structural changes of the epididymal epithelial cells were observed by transmission electron microscope.AnnexinV-FITC/PI staining was used to quantify the percentages of apoptosis in the total cell population.The TdTmediated dUTP nick end labeling(TUNEL) technique was applied to observe the morphological changes of apoptotic cells.The expression of tumor necrosis factor alpha(TNF-α) mRNA was investigated by real-time RT-PCR.The level of TNF-α in cell culture supernatant was measured by enzyme-linked immunosorbent assay(ELISA) technology.Western blot was per-formed to determine the protein expression of cytochrome C, caspase-3, caspase-8, caspase-9.Results Podophyllotoxin significantly inhibited the activity of proliferation and induced apoptosis of epididymal epithelial cells in a dose-and time-dependent manner(P<0.05), with a 50％ inhibitory concentration(IC50) value and corresponding 95％ confidence intervals(CI) of 59.36(15.50～227.41), 0.37(0.080～1.70), 0.077(0.017～0.35) μmol·L-1 at 24, 48 and 72 h, respectively.Podophyllotoxin induced cell volume turned round and cell nuclear fragmentation and mitochondrial vacuolation.RT-PCR and ELISA results showed that podophyllotoxin improved the expression of TNF-α mRNA and protein.Western blot results demonstrated that podophyllotoxin activated the protein expression of cytochrome C, caspase-3, caspase-8 and caspase-9.Conclusion Podophyllotoxin can induce rat epididymal epithelial cell apoptosis through both the mitochondria-regulated intrinsic pathway and the TNF receptor-mediated extrinsic pathway.

Objective To study the physicochemical properties of aqueous solution of surface active drug montelukast so-dium (MS) ,which could provide experimental basis for further development of micelle or mixed micelle preparations .Methods Critical micelle concentration (CMC) of MS at different temperatures were determined by conductivity measurements .The absorbance and transmittance of MS aqueous solution were measured by UV at different sodium chloride concentration levels . The micelle stability was evaluated via high speed centrifugal .Results The CMC of MS aqueous solution at 25℃ ,30℃ ,35℃were 0 .75 ,0 .82 ,0 .90 mmol/L .The absorbance and transmittance of MS aqueous solution were affected by the sodium chlo-ride concentration and the concentration of MS itself .It was observed that a clear solution was obtained when MS concentration>7 .5 mmol/L and no precipitation was noticed even after high speed centrifugal .Conclusion Montelukast sodium is a surface active drug .Its solubility is related to MS concentration .The solubility is also sensitive to the temperature and the electrolyte concentration .These unique physicochemical properties could be used to develop micelle or mixed micelle pharmaceutical prepa-rations .

Objective:To construct the evaluation index system of core competence of specialized gynecological nurses, and to provide reference for the curriculum setting and effect evaluation of standardized training for specialized gynecological nurses.Methods:The draft evaluation index system was prepared through literature research and group discussion. From May to June 2021, two rounds of Delphi letter consultation was used among 18 experts, to complete the selection and modification of evaluation index, and to determine the index weight.Results:The effective response rate of the two rounds of Delphi expert consultation was 100%, the expert authority coefficient was 0.938, and the Kendall harmony coefficient of the two rounds of expert letter consultation was 0.117 and 0.304, respectively (both P<0.01). Finally, the evaluation index system of core competence of specialized gynecological nurses was formed, including 4 first-level indexes, 11 second-level indexes and 48 third-level indexes. Conclusions:The evaluation index system of core competence of specialized gynecological nurses established in this study is scientific and reliable, which can provide reference for the curriculum setting and effect evaluation of standardized training for specialized gynecological nurses.

Objective:To explore the clinical characteristics and molecular basis for children and adolescents with monogenic diabetes.Methods:A retrospective analysis was carried out for the clinical manifestations and laboratory data of 116 children and adolescents diagnosed with diabetes at Ningbo Women and Children′s Hospital from January 2020 to March 2023. Whole exome sequencing and mitochondrial gene sequencing were carried out on 21 children with suspected monogenic diabetes.Results:A total of 10 cases of monogenic diabetes were diagnosed, all of which were Maturity-onset Diabetes Of the Young (MODY). Six cases of MODY2 were due to GCK gene mutations, 1 case of MODY3 was due to HNF1A gene mutation, 2 cases of MODY12 were due to ABCC8 gene mutations, and 1 case of MODY13 was due to KCNJ11 gene mutation. Nine of the 10 patients with MODY had no typical symptoms of diabetes. A family history of diabetes was significantly more common in the MODY group compared with the T1DM and T2DM groups ( P<0.05). The BMI of the MODY group was higher than that of the T1DM group ( P<0.05). The initial blood glucose level was lower than that of the T1DM group ( P<0.05), and there was no significant difference compared with the T2DM group. The fasting C-peptide level of the MODY group was higher than that of the T1DM group ( P<0.05), and there was no significant difference compared with the T2DM group. Glycosylated hemoglobin of the MODY group was lower than both the T1DM and T2DM groups ( P<0.05). Conclusion:In this study, MODY has accounted for the majority of monogenic diabetes among children and adolescents, and the common mutations were those of the GCK gene in association with MODY2. Blood glucose and glycosylated hemoglobin of children with MODY were slightly increased, whilst the islet cell function had remained, and the clinical manifestations and laboratory tests had overlapped with those of type 2 diabetes. WES and mitochondrial gene sequencing can clarify the etiology of monogenic diabetes and facilitate precise treatment.